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作 者:李娜 李涛[1] 杨冬梨 姚媛 LI Na;LI Tao;YANG Dongli;YAO Yuan(College of Basic Medicine,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Second Clinical Medical College,Shaanxi University of Chinese Medicine,Xianyang 712046,China)
机构地区:[1]陕西中医药大学基础医学院,陕西咸阳712046 [2]陕西中医药大学第二临床医学院,陕西咸阳712046
出 处:《基础医学与临床》2024年第4期483-488,共6页Basic and Clinical Medicine
基 金:陕西省自然科学基金(2022JQ-790,2023-JC-QN-0852);国家自然科学基金(82205030);陕西中医药大学博士启动经费(17102032006);陕西中医药大学大创项目(202210716113)。
摘 要:目的 研究厚朴酚对人脂肪间充质干细胞(hADSCs)增殖和凋亡的影响,以此细胞模型初探该药物对肿瘤微环境的影响。方法 用不同浓度的和厚朴酚处理hADSCs,分别采用MTS法和台盼蓝染色法检测细胞的增殖能力,annexin V/PI双染法检测细胞凋亡的改变,qPCR和Western blot检测细胞增殖、凋亡相关基因的mRNA和蛋白质表达,Western blot检测MEK-ERK1/2信号通路中总MEK、磷酸化MEK、总ERK和磷酸化ERK蛋白的表达水平。结果 随着浓度增加,和厚朴酚抑制hADSCs增殖、促进其凋亡的作用显著增强。增殖相关基因CCND1、MKI67和PCNA表达下调,促凋亡相关基因BAX和TP53表达上调,抗凋亡基因BCL2表达下调。和厚朴酚呈浓度依赖性抑制MEK和ERK1/2的磷酸化。结论 和厚朴酚抑制hADSCs增殖,促进其凋亡,作用机制可能与抑制MEK-ERK1/2通路活性有关。Objective To investigate the effects of honokiol on proliferation and apoptosis of human adipose-derived mesenchymal stem cells(hADSCs),and to investigate the effect of the drug on the tumor microenvironment.Methods hADSCs were incubated with different concentrations of honokiol,the proliferation of hADSCs was detected by MTS and Trypan blue staining,and cell apoptosis was assessed by annexin V/PI double staining.In the meantime,expression of mRNA and protein related to cell proliferation and apoptosis were detected by qPCR and Western blot,respectively.The expression of total MEK,phosphorylated MEK,total ERK and phosphorylated ERK proteins in the MEK-ERK1/2 signaling pathway were detected by Western blot.Results The effect of honokiol on inhibiting proliferation and promoting apoptosis of hADSCs was significantly enhanced with the increase of concentration.The expressions of proliferation-related genes CCND1,MKI67 and PCNA were down-regulated.The expressions of pro-apoptotic genes BAX and TP53 was up-regulated,and the expressions of anti-apoptotic gene BCL2 was down-regulated.Honokiol inhibited MEK and ERK1/2 phosphorylation in a concentration-dependent manner.Conclusions Honokiol inhibits proliferation and promotes apoptosis of hADSCs,and the specific mechanism is potentially related to the inhibition of MEK-ERK1/2 pathway.
关 键 词:和厚朴酚 人脂肪间充质干细胞 细胞增殖 细胞凋亡 MEK-ERK1/2
分 类 号:R329.28[医药卫生—人体解剖和组织胚胎学]
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